Abstract
A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 20 (MMP-20, enamelysin) was established by use of a solid-phase monoclonal antibody and a separate peroxidase-labeled monoclonal antibody. The EIA system was shown to be sensitive and quantitative for the detection of MMP-20. As little as 1.0 ng/ml (50 pg/assay) of MMP-20 protein could be reliably detected. The EIA system was linear over a range of 2.5-160 ng/ml (125-8,000 pg/assay), and the EIA system was versatile in that it was capable of detecting with equal sensitivity proMMP-20, active MMP-20, and MMP-20 with COOH-terminal deletions. The EIA system was validated by the successful detection of MMP-20 in the culture medium of Chinese hamster ovary cells (CHO-K1) that were transfected with an MMP-20 expression vector. No MMP-20 was detected in normal human serum, normal saliva, or in selected tumors. However, when recombinant human MMP-20 was added to human saliva, the EIA system did detect quantifiable amounts of the MMP-20, indicating that the system will work within the framework of complex in biological fluids.
| Original language | English |
|---|---|
| Pages (from-to) | 530-537 |
| Number of pages | 8 |
| Journal | European Journal of Oral Sciences |
| Volume | 108 |
| Issue number | 6 |
| Publication status | Published - 2000 Dec |
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Keywords
- Enamelysin
- MMP-20
- Monoclonal antibody
- Sandwich enzyme immunoassay
ASJC Scopus subject areas
- Dentistry(all)
Cite this
One-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human enamelysin (MMP-20). / Wang, Ting; Aoki, Takanori; Iwata, Kazushi; Takata, Takashi; Uchida, Takashi; Knäuper, Vera; Llano, Elena; Okada, Yasunori; Bartlett, John D.
In: European Journal of Oral Sciences, Vol. 108, No. 6, 12.2000, p. 530-537.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - One-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human enamelysin (MMP-20)
AU - Wang, Ting
AU - Aoki, Takanori
AU - Iwata, Kazushi
AU - Takata, Takashi
AU - Uchida, Takashi
AU - Knäuper, Vera
AU - Llano, Elena
AU - Okada, Yasunori
AU - Bartlett, John D.
PY - 2000/12
Y1 - 2000/12
N2 - A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 20 (MMP-20, enamelysin) was established by use of a solid-phase monoclonal antibody and a separate peroxidase-labeled monoclonal antibody. The EIA system was shown to be sensitive and quantitative for the detection of MMP-20. As little as 1.0 ng/ml (50 pg/assay) of MMP-20 protein could be reliably detected. The EIA system was linear over a range of 2.5-160 ng/ml (125-8,000 pg/assay), and the EIA system was versatile in that it was capable of detecting with equal sensitivity proMMP-20, active MMP-20, and MMP-20 with COOH-terminal deletions. The EIA system was validated by the successful detection of MMP-20 in the culture medium of Chinese hamster ovary cells (CHO-K1) that were transfected with an MMP-20 expression vector. No MMP-20 was detected in normal human serum, normal saliva, or in selected tumors. However, when recombinant human MMP-20 was added to human saliva, the EIA system did detect quantifiable amounts of the MMP-20, indicating that the system will work within the framework of complex in biological fluids.
AB - A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 20 (MMP-20, enamelysin) was established by use of a solid-phase monoclonal antibody and a separate peroxidase-labeled monoclonal antibody. The EIA system was shown to be sensitive and quantitative for the detection of MMP-20. As little as 1.0 ng/ml (50 pg/assay) of MMP-20 protein could be reliably detected. The EIA system was linear over a range of 2.5-160 ng/ml (125-8,000 pg/assay), and the EIA system was versatile in that it was capable of detecting with equal sensitivity proMMP-20, active MMP-20, and MMP-20 with COOH-terminal deletions. The EIA system was validated by the successful detection of MMP-20 in the culture medium of Chinese hamster ovary cells (CHO-K1) that were transfected with an MMP-20 expression vector. No MMP-20 was detected in normal human serum, normal saliva, or in selected tumors. However, when recombinant human MMP-20 was added to human saliva, the EIA system did detect quantifiable amounts of the MMP-20, indicating that the system will work within the framework of complex in biological fluids.
KW - Enamelysin
KW - MMP-20
KW - Monoclonal antibody
KW - Sandwich enzyme immunoassay
UR - http://www.scopus.com/inward/record.url?scp=0034567330&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034567330&partnerID=8YFLogxK
M3 - Article
VL - 108
SP - 530
EP - 537
JO - European Journal of Oral Sciences
JF - European Journal of Oral Sciences
SN - 0909-8836
IS - 6
ER -